Medicinal product for the promotion of wound healing

ABSTRACT

The invention relates to a medicinal product for topical use for the promotion of wound healing, which comprises thrombocytes or thrombocyte fragments, wherein the thrombocytes or thrombocyte fragments contain growth factors and are capable of releasing the same and are present in the lyophilized or deep-frozen state and have been subjected to a process for virus partitioning and/or virus inactivation.

[0001] This invention relates to a medicinal product for topical use forthe promotion of wound healing.

[0002] It is known that the healing of a wound progresses in severalsuccessive stages.

[0003] In stage I, the blood plasma protein fibrinogen is precipitatedby thrombin so as to induce the formation of a fibrin clot, whichsolidifies in the presence of blood coagulation factor XIII. In thefirst stage which takes only minutes bleeding is controlled and thewound area is sealed.

[0004] In stage II, cells from the wound area migrate into the fibrinclot, i.e., inflammatory cells, connective tissue cells and endothelialcells. They form vessels and, as an extracellular matrix, connectivetissue primarily comprised of collagen. This connective tissue, which isreferred to as granulation tissue, serves as the substratum for theformation of epithelial tissue and is the substratum for the epidermison the body surface. Stage II lasts for days to weeks and is complete assoon as the wound area has been closed by epithelium, and by theepidermis on the skin.

[0005] Wound healing is complete by stage II, which lasts for weeks tomonths. During that phase, the cellular elements are reduced and theconnective tissue is growing so as to form a firm and permanent scartissue. (Bennett N. T., Schultz G. S., Am. J. Surg. 1993, 165:728-737;Bennett N. T., Schultz G. S., Am. J. Surg. 1993, 166: 74-81).

[0006] The formation of granulation tissue in stage II of the woundhealing process is effected by growth factors promoting the migrationand the division of connective tissue cells as well as the regenerationof vessels and, thereby, accelerating wound healing. Of the known growthfactors, platelet derived growth factor (PDGF), transforming growthfactor β (TGF-β), epidermal growth factor (EGF) and insulin-like growthfactor I (IGF-I) are particularly involved in those processes. (BennettN. T., Schultz G. S., Am. J. Surg. 1993, 165: 728-737; Bennett N. T.,Schultz G. S., Am. J. Surg. 1993, 166: 74-81; Bhora F. Y. et al., J.Surg. Res. 1995, 59: 236-244; Lynch S. E. et al., Proc. Natl. Acad. Sci.USA 1987, 84: 640-646; Lynch S. E. et al., J. Clin. Invest. 1989, 84:7696-7700).

[0007] Also the regeneration of the epidermis is induced by growthfactors. They activate the epidermal cells (keratinocytes) that havebeen detached from the cell association of the intact basal cell layerdue to the lesion, so as to form specific membrane receptors enablingthe adherence to the granulation tissue substratum, in particular tofibrin-fibronectin, which constitutes a provisional scaffold forkeratinocyte migration (Brown G. L. et al., J. Exp. Med. 1986, 163:1319-1324; Brown G. L. et al., N. Engl. J. Med. 1989, 321: 76-79).

[0008] Growth factors are synthesized in the human body by varioustissues and cell types and secreted into the surrounding body liquid. Inthe context of wound healing, an important regulatory role is attributedto thrombocytes, which are able to synthesize in significant amounts andstore growth factors PDGF, TGF-β, EGF and IGF-I, which are essential towound healing in cytoplasmic granula. (Lynch S. E. et al., Proc. Natl.Acad. Sci. USA 1987; 84: 640-646; Ginsberg M. H. et al., Thromb.Haemostas. 1988, 59: 1-6; Hyner O. R., Thromb. Haemostas. 1991, 66:40-43).

[0009] In order to release or deliver the stored growth factors from thethrombocytes, the latter must be activated by physiological stimuli suchas, e.g., collagen, thrombin, trypsin, ADP, serotonin or adrenalin,which bind to specific receptors on the external surface of thethrombocyte plasma membrane. Activation results in a change of shapefollowed by the aggregation of thrombocytes, whereupon the lattersecrete the stored growth factors into the surrounding body liquid. Withmost of these physiological stimuli, the aggregation of thrombocytesfollowing activation is a prerequisite for the release of growthfactors. By stimulation with thrombin, growth factors may be releasedalso without thrombocyte aggregation. (Kaplan K. L. et al., Blood 1979,53: 604-618; Holmsen H. et al., J. Biol. Chem. 1981, 256: 9393-9396;Philipps D. R., Baughan A. K., J. Biol. Chem. 1983, 258: 10240-10245).

[0010] The interactions between activated thrombocytes, which lead toaggregation, and their adherence to surfaces are mediated byextracellular adhesive matrix proteins such as, e.g., fibrinogen,fibronectin and von Willebrand factor, which bind to a glycoproteinreceptor on the external side of the plasma membrane of the activatedthrombocytes. Strong binding of these matrix proteins to the receptor iseffected only where thrombocytes have been activated by an appropriatestimulus as described above. These complex procedures of thrombocyteactivation and aggregation followed by the release of growth factorsconstitute one of the essential control elements in the wound healingprocess. (Ginsberg M. H. et al., Thromb. Haemostas. 1988, 59: 1-6; HynerO. R., Thromb. Haemostas. 1991, 66: 40-43; Landolfi R. et al., Blood1991, 78: 377-381; Perschke E. I. et al., Blood 1980, 55: 841-847; HynesO. R., Cell 1992, 69: 11-25; Perschke E. I., J. Lab. Clin. Med. 1994,124: 439-446; Savage B.,Ruggeri Z. M., J. Biol. Chem. 1991, 266:11227-11233; Bennett J. S. et al., J. Biol. Chem. 1982, 257: 8049-8054;Cierniewski C. S. et al., Biochim. Biophys. Acta 1982, 714: 543-548;Philipps D. R., Baughan A. K., J. Biol. Chem. 1983, 258: 10240-10245).

[0011] Disturbances in wound healing as these occur, for instance, inpatients with diabetes, venous or arterial occlusions, but also woundhealing disturbances of other geneses such as, for instance, irradiationwith radioactive substances or after bums particularly affect stage IIof the wound healing process. It has been found that in such casesgrowth factors are present to a reduced extent so that no or only a lowquality granulation tissue is formed. (Dvonch V. M. et al., Surgery1992, 112: 18-23; Matsuoka J., Grotendorst G. R., Proc. Natl. Acad. Sci.USA 1989, 86: 4416-4420).

[0012] In order to enhance wound healing in the case of wound healingdisturbances, growth factors are known to be applied to the wound area,either individually or in combination, as a pure substance or mixed inointment bases (Knighton D. R. et al., Surg. Gynecol. Obstet. 1990, 170:56-60; Brown G. L. et al., J. Exp. Med. 1986,163: 1319-1324; Holmsen H.et al., J. Biol. Chem. 1981, 256: 9393-9396). The growth factorsprovided in this manner are, however, rapidly inactivated or degradedand develop their activities only over short periods of time (minutes)after application. Thus, these preparations offer no satisfactoryenhancement of wound healing.

[0013] Other known therapeutic approaches consist in covering the woundarea with collagen sponges or other preparations aimed to ensurepermanent humidity of the wound area or in using preparations degradingthe superficial connective tissue layer of the wound area byfermentation so as to enable new connective tissue to re-grow from thewound bed (Nielsen P. G. et al., Acta Dermato-Venerologica 1990, Suppl.152: 1-12; Lippert P., Wolff H., Zent.bl. Chir. 1990, 115: 1175-1180).Yet, none of those hitherto applied wound dressings and preparations ormedicinal products have brought satisfactory results in improving woundhealing.

[0014] The object of the present invention is to provide a medicinalproduct which efficaciously accelerates natural wound healing processesand is capable of substantially improving wound healing where woundhealing is disturbed, in particular in severe forms of wound healingdisturbances, as compared to conventional therapies.

[0015] In accordance with the invention, this object is achieved in thata medicinal product for topical use for the promotion of wound healingis provided, which comprises thrombocytes or thrombocyte fragments,wherein said thrombocytes or thrombocyte fragments contain growthfactors and are capable of releasing the same, are present in thelyophilized or deep-frozen state and have been subjected to a processfor virus partitioning and/or virus inactivation.

[0016] “Thrombocyte fragments” is intended to denote any insolublethrombocyte constituents that are separable from the soluble thrombocyteconstituents either by filtration including nano-filtration or bycentrifugation including ultracentrifugation.

[0017] Unless indicated otherwise, the term “thrombocytes” in thefollowing also encompasses “thrombocyte fragments”.

[0018] The invention is based on the finding that the topical use ofthrombocytes containing growth factors and capable of releasing the samecan efficaciously accelerate wound healing processes. The thrombocytesapplied on the wound area constitute a natural reservoir for the growthfactors required for the promotion of the wound healing processes. Ithas been found that the activation of locally applied thrombocytes byphysiological stimuli present in the wound area and the subsequentaggregation and binding of the matrix proteins present in the wound arealead the growth factors stored in the thrombocytes to be released intothe wound area continuously over an extended period of time (severaldays). Due to this fact, higher concentrations of growth factors areapparently availabe in the wound area over a substantially longer periodof time than with the direct administration of growth factors, therebypromoting the immigration of inflammatory cells, connective tissue cellsand endothelial cells and enhancing the propagation of said cells instage II of the wound healing process. In that manner, the rapid andsufficient formation of granulation tissue is ensured, which, in turn,renders possible the formation of epithelial tissue and the final woundclosure. The epithelization process, moreover, is additionallyaccelerated by the released growth factors promoting the immigration andproliferation of epithelial cells.

[0019] To ensure that the medicinal product can be stored over anextended period of time, the thrombocytes in the medicinal productaccording to the invention preferably are present in the lyophilized ordeep-frozen state. In order to minimize the risk of virus infections,the thrombocytes advantageously are subjected to a process for viruspartitioning and/or virus inactivation, whereby a physical or a chemicalor a combined process may be used.

[0020] In order to provide for a higher concentration of growth factors,in particular in the treatment of wound healing disturbances, it ispreferred that the content of thrombocytes or thrombocyte fragments ofthe medicinal product according to the invention is such that itcorresponds to at least 10⁴, preferably at least 10⁵, thrombocytes perμl after reconstitution of the lyophilisate or thawing.

[0021] In order to obtain a particularly pronounced initial effect ofthe medicinal product according to the invention immediately uponapplication, it may be appropriate, in particular in the case of severedisturbances of wound healing, that the medicinal product comprisesadditional growth factors that are not derived from the thrombocytescontained in the medicinal product. The additional growth factors may beof the same type as those stored and released by the thrombocytes of themedicinal product according to the invention or belong to a differenttype. The growth factors may be present in the same container with thethrombocytes or contained in a separate container in the form of asolution or lyophilisate.

[0022] It has been found that it is advantageous, in particular insevere cases of disturbed wound healing, that the medicinal productcomprises biomaterials. “Biomaterials” in the sense of the invention isintended to comprise any materials which are tissue-compatible andabsorbable and assist in the promotion of wound healing either incombination with the thrombocytes or growth factors contained in themedicinal product or independently thereof. Thus, substances activatingthrombocytes as stimuli and/or materials mediating thrombocyteaggregation may be contained as biomaterials in the medicinal productaccording to the invention. In that manner, the activity of naturalsubstances present in the wound area which activate thrombocytes andmediate their aggregation is enhanced, which increases the release ofgrowth factors and promotes wound healing even further.

[0023] In order to minimize the risk of virus infections, thebiomaterials preferably are subjected to a process for viruspartitioning and/or virus inactivation, wherein a physical or chemicalprocess or a combined process may be applied. The biomaterials may besubjected to such a process either individually or mixed with othercomponents of the medicinal product (e.g., thrombocytes).

[0024] To ensure that the medicinal product can be stored over anextended period of time, the biomaterials in the medicinal productaccording to the invention advantageously are present in the lyophilizedor deep-frozen state. In that case, the biomaterials may be present inthe same containers with the thrombocytes and/or growth factors orcontained in separate containers and deep-freezing or lyophilization ofthe biomaterials may be effected individually or in mixture with othercomponents of the medicinal product.

[0025] It is known that the activation and aggregation of thrombocytesand hence the release of growth factors stored in the thrombocytes isenabled by the attachment of matrix proteins. Moreover, such proteinsmay form cross-linked structures to which the thrombocytes adhere andfirmly bind to the wound area, such structures promoting the diffusionof growth factors to the wound area and the immigration of cells fromthe wound area. Accordingly, a preferred embodiment of the medicinalproduct according to the invention is characterized in that tissueadhesive and/or collagen are provided as biomaterials. Tissue adhesivein the sense of the invention is intended to encompass biomaterialstotally or partially consisting of cross-linkable proteins suitable fortissue adhesion.

[0026] Fibrinogen is a particularly active substance for triggering theaggregation of activated thrombocytes, while thrombin represents one ofthe most active substances for the activation of thrombocytes. It is,therefore, advantageous for an increase in the relase of growth factorsand an enhancement of wound healing that the tissue adhesive is composedof fibrinogen-containing proteins and thrombin.

[0027] It has been shown that human cells such as keratinocytes,epithelial cells, embryonic and fetal cells as well as cell constituentssuch as liposomes are able to additionally acceleratethrombocyte-promoted wound healing and cell propagation. It is,therefore, preferred that the medicinal product additionally comprisesepithelial cells and/or keratinocytes and/or embryonic and/or fetalcells and/or liposomes. The cells or the liposomes may be present as aliquid or deep-frozen suspension or as a lyophilisate in separatecontainers, or one or several of the mentioned cell types or liposomeseither without or with any of the other components of the medicinalproduct in common containers.

[0028] In order to minimize the risk of virus infections, the cells orthe liposomes may have been subjected to a process for viruspartitioning and/or virus inactivation, whereby a physical or a chemicalprocess or a combined process may be used. The cells or the liposomesmay be subjected to such a process either individually or mixed withother components of the medicinal product.

[0029] The invention also relates to the use of thrombocytes orthrombocyte fragments containing growth factors for the production of amedicinal product for topical use for the promotion of wound healing.

[0030] Preferred embodiments of the invention will now be explained inmore detail by way of examples.

EXAMPLE 1

[0031] Preparation of a Medicinal Product According to the Invention

[0032] A human thrombocyte concentrate or concentrate of thrombocyteconstituents is anticoagulated by 3% sodium citrate and centrifuged(1000 g/20 min) in order to eliminate plasma and other cellconstituents. The thrombocyte-rich supernatant, or supernatant ofthrombocyte constituents, is suspended in RPMI medium and washed threetimes in RPMI medium (1000 g/20 min). The washed thrombocytes, or thewashed thrombocyte constituents, are suspended in RPMI medium andadjusted to a concentration of at least 6×10⁵ thrombocytes orthrombocyte constituents per μl. After this, the thrombocyte suspensionis subjected to a virus inactivation process according to Example 3 andsubsequently deep-frozen or lyophilized in accordance with the methodsdescribed below, thereby obtaining a medicinal product according to theinvention.

[0033] Deep-freezing:

[0034] 1 ml of the thrombocyte suspension is each shock deep-frozen at−80° C. within 30-40 minutes and stored in a deep-frozen state. Beforeuse, the thrombocyte concentrate is thawed at room temperature.

[0035] Lyophilization:

[0036] 1 ml of the thrombocyte suspension is each shock deep-frozen at−80° C. for at least 24 hours and subsequently lyophilized at −20° C. to−40° C. in vacuo for 20 to 24 hours. The lyophilized thrombocytes arestored at between −20° C. and −80° C. and rehydrated with 1 ml RPMImedium before use.

EXAMPLE 2

[0037] Preparation of a Medicinal Product According to the InventionComprising Biomaterials

[0038] The virus-inactivated thrombocyte suspension prepared accordingto Example 1 is supplemented with a solution of cross-linkable humanprotein (either fibrinogen, fibronectin, blood coagulation factor XIIIor collagen) which may have been subjected to one or several processesfor virus inactivation according to Example 4, each protein typeseparately or together in combination, wherein the concentration of thecross-linkable protein types in the supplemented solution preferably isto amount to 70-90 mg/ml. The mixing ratio of the thrombocyte suspensionto the solution of cross-linkable human protein preferably is to be 1:3.The thus obtained mixture is deep-frozen or lyophilized in accordancewith the processes described in Example 1 in order to obtain suitablestorability.

[0039] Instead of carrying out virus inactivation on individualcomponents (thrombocytes or biomaterials), it is also possible to effectvirus inactivation on a mixture of thrombocyte suspension and proteinsolution according to the process of Example 3.

EXAMPLE 3

[0040] Virus Inactivation of Thrombocyte Suspension (Photodynamic VirusInactivation)

[0041] To 50 ml of the thrombocyte suspension prepared according toExample 1 is added 8-methoxypsoralen (dissolved in dimethylsulfoxide[DMSO]) until a final concentration of 300 μl/ml (final concentration ofDMSO 0.3%) and irradiated with ultraviolet light from below and abovefor 6 hours at 22-27° C. under an atmosphere of 5% CO₂ and 95% N₂ and ata pressure of 2 psi such that the overall light intensity is 3.5 to 4.8mW/cm² (Lin L. et al., Blood 1989, 74: 517-525).

[0042] After photoinactivation has been completed, the thrombocytesuspensions obtained in that manner are examined for their finctionalcapacities. The functional capacity is determined by measuring[³H]-thymidine incorporation in a fibroblast cell culture.

EXAMPLE 4

[0043] Virus Inactivation of Biomaterials (Chemical Virus Inactivation)

[0044] Biomaterials which are admixed to the thrombocyte suspensionprepared according to Example 1 are virus inactivated by a solventdetergent method. To this end, a biomaterial suspension is supplementedwith 1% (w/w) tri(n-butyl) phosphate and 1% (w/w) Triton X-100 at 30° C.and the mixture is kept for 4 hours under shaking. After this, thesolvent detergent mixture under the addition of 5% (v/v) soybean oil isremoved from the biomaterial suspension by chromatography on aC18-column (Waters Millipore) (Horowitz B. et al., Blood 1992, 79:826-831; Piet M. P. J. et al., Transfusion 1990, 30:591-598; Piquet Y.et al., Vox sang. 1992, 63: 251-256).

[0045] The biomaterials treated by the above-described chemical virusinactivation method may subsequently be subjected to photodynamic virusinactivation in addition.

EXAMPLE 5

[0046] Evaluation of the Promotion of Connective Tissue Proliferation bythe Medicinal Product According to the Invention

[0047] The test was carried out on a fibroblast cell culture. Themedicinal product prepared according to Example 2 was applied on a cellculture plate in an amount of 200 μl per cm² and activated by 50 μl of athrombin solution (3.2 IU thrombin per ml physiological saline). Humanfibroblasts derived from the 4^(th) to 10^(th) passages of a primaryculture were placed on the applied suspension at a density of 4×10⁴cells per cm² and cultivated in cell culture medium (RPMI) (culture 1).On the third, fifth and seventh days of cultivation, the cell mitoticrate was measured by measuring DNA synthesis via [³H]-thymidineincorporation. The cell mitotic rate of culture 1 was compared to thecell mitotic rate of another fibroblast culture (culture 2) realized inRPMI nutrient supplemented with 10% by vol. of calf serum withoutaddition of the medicinal product according to the invention.

[0048] Results:

[0049] On day 3 of cultivation, culture 1 exhibited a [³H]-thymidineincorporation (196645±56864 cpm/ml) that was seven times higher thanthat of culture 2. On days 5 (152749±93951 cpm/ml) and 7 (77045±27974cpm/ml) [³H]-thymidine incorporation in culture 1 still was 5 to 10times higher than that of culture 2. These differences between culture 1and culture 2 statistically are highly significant (p<0.01),demonstrating the ability of the medicinal product according to theinvention to promote connective tissue proliferation and maintain thatactivity over an extended period of time (at least 7 days).

EXAMPLE 6

[0050] Evaluation of the Binding of Matrix Proteins to ThrombocyteSurfaces Resulting in the Thrombocyte Stored Growth Factors to beContinuously Released

[0051] The test was carried out on a fibroblast culture (according toExample 5). Culture 1—as in Example 5—was supplemented with themedicinal product according to the invention. In culture 2, thethrombocytes were treated with specific antibodies, against thesuperficial binding sites for matrix proteins so as to prevent thematrix proteins from binding to thrombocyte surfaces. On the third dayof cultivation, the cell mitotic rate was measured by measuring DNAsynthesis via [³H]-thymidine incorporation.

[0052] Results:

[0053] While culture 1 exhibited a thymidine incorporation rate similarto that of Example 5, no thymidine incorporation could be measured inculture 2. That difference proves that the binding of matrix proteins tothe thrombocyte surfaces is necessary for the thrombocyte stored growthfactors to be released.

EXAMPLE 7

[0054] Evaluation of the Promotion of Wound Healing by the MedicinalProduct According to the Invention

[0055] The clinical efficacy of the medicinal product according to theinvention was studied in six patients suffering from chronic,non-healing cutaneous ulcera of the lower extremities and alreadytreated by surgical or conservative topical therapies for more than sixmonths without success. The ulcera were classified using a wound scoreaccording to Knighton D. R. et al., Ann. Surg. 1986, 204:322-330. Thewound score includes general parameters, anatomical conditions andmeasurable variables of the ulcus. The higher the scores, the poorer thepreconditions for healing; the highest score to be reached is 97 (=worststarting situation).

[0056] Treatment Plan:

[0057] The ulcera were cleaned, necrotic tissue was removed and wettedwith a thrombin solution (3.2 IU bovine thrombin/ml RPMI medium). Afterthis, the defect was filled up with the thawed medicinal productaccording to the invention prepared according to Example 2, and theabove-mentioned thrombin solution was then applied at a volume ratio ofmedicinal product suspension to thrombin solution of 3:1 in order toactivate the thrombocytes. The ulcera eated in that manner were coveredby a non-adhering wound dressing (metal foil). Until healing, the ulcerawere treated twice a week in the above-identified manner. The healingprogress was documented photographically and histologically (fine needlebiopsies in the 2^(nd) and 5^(th) weeks of treatment).

[0058] Relults:

[0059] The demographics, causative vascular and metabolic diseases ofthe patients and the evaluation of the wound scores at the beginning oftreatment are summarized in Table 1. TABLE 1 Vascular Disease MetabolicWound Patient Sex Age arterial venous disease Score 1 male 67 + +diabetes 51 2 male 72 + − — 65 3 male 69 + − diabetes 33 4 male 63 + −diabetes 49 5 male 78 + + diabetes 63 6 female 74 − + — 65^(a)/63^(b)

[0060] The time course of wound healing (indicated in weeks as of thebeginning of treatment) is illustrated in Table 2. TABLE 2 Beginning ofGranulation Tissue Beginning of Completion of Patient FormationEpithelization Epithelization 1 1st week 3rd week 8th week 2 1st week3rd week 9th week 3 3rd week 8th week 12th week 4 1st week 4th week 10thweek 5 1st week none none 6 ^(a,b)1st week ^(a)6th/^(b)3rd week^(a)12th/^(b)9th week

[0061] With the exception of patient 3, a granulation tissue wellsupplied with blood formed starting from the bottom of the ulcus in allof the patients already within the first week of treatment, whichgranulation tissue increased upon further treatment with the medicinalproduct according to the invention until approximately two weeks afterthe beginning of the therapy and filled up the ulcus. It was strikingthat already after the first days of treatment the surrounding of theulcus calmed down, the erythema and the edema of the surrounding skindisappeared and also the edge of the ulcus was no longer edematous andmiscolored in all of the patients. Histologically, cell-rich granulationtissue primarily consisting of fibroblasts and fibrocytes and exhibitingintensive new vascular formation and collagenous fiber formation andonly a slight infiltration of inflammatory cells and tissue necroses onthe surface was to be seen in all biopsies in the second week oftreatment. Epithelization of the skin defects after the third week oftreatment started from the edges of the wound and could then also bedetected histologically by the second biopsies in the fifth week oftreatment. In the further course of treatment, the size of the ulceradeclined due to epithelization, but also to cicatricial shrinkage. Withthe exception of patient 5, they were scarred over in the 12^(th) weekof treatment the latest.

[0062] The results indicated above demonstrate that the topical use ofthe medicinal product according to the invention promotes wound healingand, thus, is able to completely cure chemically non-healing cutaneousulcera in patients treated by conservative therapies for at least sixmonths without success and, thus, offering extremely poor prognoses forwound healing.

1. A medicinal product for topical use for the promotion of woundhealing, which comprises thrombocytes or thrombocyte fragments, whereinsaid thrombocytes or thrombocyte fragments contain growth factors andare capable of releasing the same, are present in the lyophilized ordeep-frozen state, and have been subjected to a process for viruspartitioning and/or virus inactivation.
 2. A medicinal product accordingto claim 1, characterized in that the content of thrombocytes orthrombocyte fragments is such that it corresponds to at least 10⁴,preferably at least 10⁵, thrombocytes per ml after reconstitution of thelyophilisate or thawing.
 3. A medicinal product according to claim 1 or2, characterized in that the medicinal product comprises additionalgrowth factors.
 4. A medicinal product according to any one of claims 1to 3, characterized in that the medicinal product comprisesbiomaterials.
 5. A medicinal product according to claim 4, characterizedin that the biomaterials have been subjected to a process for viruspartitioning and/or virus inactivation.
 6. A medicinal product accordingto claim 4 or 5, characterized in that the biomaterials are present inthe lyophilized or deep-frozen state.
 7. A medicinal product accordingto any one of claims 4 to 6, characterized in that tissue adhesiveand/or collagen are provided as biomaterials.
 8. A medicinal productaccording to claim 7, characterized in that the tissue adhesive iscomposed of fibrinogen-containing proteins and thrombin.
 9. A medicinalproduct according to any one of claims 4 to 8, characterized in that themedicinal product additionally comprises epithelial cells and/orkeratinocytes and/or embryonic and/or fetal cells and/or liposomes. 10.The use of thrombocytes or thrombocyte fragments containing growthfactors and capable of releasing the same, for the production of amedicinal product for topical use for the promotion of wound healing.